| Objective: To construct an animal model of irritable bowel syndrome(IBS-D)with liver-depression and spleen-deficiency,to detect cAMP,PKA and IL-8 in colonic tissues by Western-bolt,to test the expression of cAMP and PKA in colonic tissues by Quantitative Real-time PCR and to detect the change of IL-8 in serum by ELISA,and to observe the histopathology of colonic tissues by HE staining.The histopathology of the colon was observed by HE staining,and the effect of an intestinal soup on the treatment of IBS-D through this pathway was analyzed,thus alleviating the visceral hypersensitivity,which provides new ideas for the clinical diagnosis and treatment of diarrheal irritable bowel syndrome at the molecular level.METHODS: Forty SPF SD male rats were selected and divided into four groups according to the random number table method,namely,normal group,blank group,an intestinal soup group and western medicine group,with 10 rats in each group.All rats were fed uniformly for 1 week,and from the beginning of the second week of the experiment,except for the normal group,the blank group,the an intestinal soup group and the western medicine group were gavaged with chorine and restrained with stress for 2 consecutive weeks to make a liver-depressed spleen-deficient IBS-D rat model,and the rats’ mental status,activity,hair,appetite,body weight,and feces,and abdominal withdrawal reflex(AWR)were observed during the modeling period.The success of the modeling was evaluated by observing the mental status,activity,hair,appetite,body weight,feces,abdominal withdrawal reflex(AWR)and abdominal wall tension.After successful modeling,the blank group was given saline 0.5mg/kg/day by gavage,the an intestinal soup group was given an intestinal soup 6.2g/kg/day by gavage,and the western medicine group was given pivibromine suspension 15mg/kg/day by gavage,and all were administered continuously for 2 weeks.The expression of cAMP,PKA and IL-8 in colonic tissues was detected by WB technique,and the expression of cAMP and PKA in colonic tissues was detected by PCR technique and the change of IL-8 in serum was detected by ELISA,and the histopathology of colonic tissues was observed by HE staining.(1)Behavior,body weight,food consumption and stool observation: the rats in the normal group had smooth fur,lively movements,good mental status,larger body weight than the modeling group,normal diet and water intake,and normal shiny stool;the rats in the modeling group had dull fur,depressed spirit,easy to hide in corners,irritable after touching,thin body weight,significantly reduced appetite and water intake compared with the normal group,and increased frequency of defecation and unformed,with The number of bowel movements increased and was unformed,with a thin and rotten stool.The behavior,body weight,food consumption and stool of the rats in the An intestinal soup group and the western medicine group were significantly improved after the intervention treatment compared with the blank group.(2)Abdominal withdrawal reflex(AWR)score of rats: the AWR score of the blank group was significantly higher than that of the normal group,Chinese medicine group and western medicine group(P<0.01),and the AWR score of the an intestinal soup group was lower than that of the western medicine group(P<0.05);(3)HE staining of rat colonic tissues: microscopically,the structure of the mucosal muscle layer of the normal group was clear,and the mucosal muscle layer of the normal group was clear.(3)HE staining of rat colon tissues: microscopically,the structure of mucosal muscle layer in the normal group was clear and no obvious pathological changes were seen;in the blank group,small focal infiltrations of lymphocytes were seen locally in the colon tissues;in the An intestinal soup group,the structure of mucosal muscle layer in the rat colon tissues was clear and no obvious pathological changes were seen;in the western medicine group,small focal infiltrations of lymphocytes were seen locally.(4)Comparison of cAMP,PKA and IL-8 in colon tissue of rats: the expression of cAMP in rats: compared with the normal group,the expression of cAMP in colon tissue of model group was significantly lower(P<0.0001),and the difference was statistically significant;compared with the model group,after treatment,the expression of cAMP in colon tissue of An intestinal soup group was higher(P<0.0001),and the difference was statistically significant The cAMP colonic tissue expression was elevated in the Western medicine group(P < 0.001),and the difference was statistically significant;after treatment,the cAMP expression in the Chinese medicine group was slightly higher than that in the Western medicine group(P < 0.05),and the difference was statistically significant;compared with the normal group,the PKA colonic tissue expression was significantly lower in the model group(P< 0.0001),and the difference was statistically significant;compared with the model group,after After treatment,PKA colonic tissue expression was elevated in the An intestinal soup group(P < 0.001),and the difference was statistically significant;PKA colonic tissue expression was elevated in the western medicine group(P < 0.01),and the difference was statistically significant;after treatment,PKA expression was slightly higher in the Chinese medicine group compared with the western medicine group(P <0.01),and the difference was statistically significant;compared with the normal group,IL-8 colonic tissue expression was Compared with the normal group,the expression of IL-8 colon tissue in the model group was significantly higher(P < 0.0001),and the difference was statistically significant;compared with the model group,after treatment,the expression of IL-8 colon tissue in the An intestinal soup group was significantly lower(P<0.001),and the difference was statistically significant,and the expression of IL-8 colon tissue in the western medicine group was significantly lower(P<0.01);after treatment,the expression of IL-8 in the traditional Chinese medicine group was slightly lower than that in the western medicine group(P<0.05),and the difference was statistically significant.0.05),and the difference was statistically significant.Conclusion: 1.It was confirmed that the cAMP,PKA and IL-8 pathways are correlated with the pathogenic process of IBS-D;2.It was confirmed that An intestinal soup can reduce intestinal hypersensitivity and improve inflammatory response by regulating the expression of cAMP and PKA and inhibiting the release of downstream inflammatory factor IL-8,thus achieving the purpose of treating IBS-D. |
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